携带GFP基因的重组狂犬病病毒的拯救及其生物学特性研究

doi:10.16431/j.cnki.1671-7236.2018.07.030

Abstract

Abstract:

The aim of this study was to construct a recombinant virus carrying green fluorescent protein (GFP) protein,and detect vaccine titer more rapidly and accurately.Recombinant plasmids containing GFP gene were constructed by gene recombination technology and rescued by reverse genetics.The viral infection in Vero cells and protein samples were collected for Western blotting assay to detect the expression of foreign protein GFP.One-step growth curve was compared with the biological characteristics of the original strain.The results showed that we successfully rescued the recombinant GFP-expressing recombinant rabies virus SAD-GFP using the constructed recombinant plasmids.Western blotting result showed that the recombinant virus could express GFP,and the expression of GFP increased with the time of virus infection.The expression of structural protein G was not affected by the expression of foreign proteins.There was no significant difference between the recombinant strains and the original strains.After passage,GFP could stably express in the virus.In this experiment,the recombinant SAD-GFP strain was successfully rescued.In the continuous passage,the GFP gene was not lost,and the protein expression level did not decrease,which proved that the foreign protein expressed at this site could sustainedly and stablly express in the course of virus passage,and did not affect the growth characteristics of the virus.

Key words: rabies virus; reverse genetic; GFP

CLC Number:

S852.65

LIU Cui, YIN Xiangping. Rescue and Biological Characteristics Analysis of Recombinant Rabies Virus Containing GFP Gene[J]. , 2018, 45(7): 1965-1971.

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Rescue and Biological Characteristics Analysis of Recombinant Rabies Virus Containing GFP Gene

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